Adenine nucleotide binding sites on beef heart F/sub 1/ ATPase: photoaffinity labeling of. beta. -subunit Tyr-368 at a noncatalytic site and. beta. Tyr-345 at a catalytic site
2-Azidoadenine (/sup 32/P)nucleotide was bound specifically at catalytic or noncatalytic nucleotide binding sites on beef heart mitochondrial F/sub 1/ ATPase. In both cases, photolysis resulted in nearly exclusive labeling of the ..beta.. subunit. The modified enzyme was digested with trypsin, and labeled peptides were purified by reversed-phase high-pressure liquid chromatography. Amino acid sequence analysis of the major /sup 32/P-labeled tryptic fragments showed ..beta..-subunit Tyr-368 to be present at noncatalytic sites and ..beta.. Tyr-345 to be present at catalytic sites. From the relationship between the degree of inhibition and extent of modification, it is estimated that one-third of the catalytic sites or two-thirds of the noncatalytic sites must be modified to give near-complete inhibition of catalytic activity.
- Research Organization:
- State Univ. of New York, Syracuse (USA)
- OSTI ID:
- 5242643
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 84:16
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ATP-ASE
CONFIGURATION INTERACTION
ENZYME ACTIVITY
PHOSPHORYLATION
NUCLEOTIDES
ADENINES
AMINO ACID SEQUENCE
BIOCHEMICAL REACTION KINETICS
CATTLE
HEART
LIQUID COLUMN CHROMATOGRAPHY
PHOSPHORUS 32
TYROSINE
ACID ANHYDRASES
AMINES
AMINO ACIDS
ANIMALS
ANTIMETABOLITES
AROMATICS
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ENZYMES
HETEROCYCLIC COMPOUNDS
HYDROLASES
HYDROXY ACIDS
ISOTOPES
KINETICS
LIGHT NUCLEI
MAMMALS
MOLECULAR STRUCTURE
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PHOSPHOHYDROLASES
PHOSPHORUS ISOTOPES
PURINES
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
SEPARATION PROCESSES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques